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Current Genetics

, Volume 36, Issue 1–2, pp 98–104 | Cite as

Transformation of the bioherbicide Colletotrichum gloeosporioides f. sp. aeschynomene by electroporation of germinated conidia

  • Micah Robinson
  • Amir Sharon
ORIGINAL PAPER

Abstract

A highly efficient and reproducible procedure for the transformation of the bioherbicide Colletotrichum gloeosporioides f. sp. aeschynomene by electroporation of germinated conidia is reported. Development of the procedure involved a detailed study of the germination process followed by extensive calibration of the electrical-pulse and selection conditions. Optimization of the transformation protocol was facilitated by the use of the green fluorescent protein that helped in the identification of stable transformants and in a fast assessment of transgene expression levels, colony homogeneity and genetic stability. Under optimal conditions, over 80 stable transformants/cuvette were obtained. Plasmid integration was predominantly homologous, but high transformation rates were obtained both with homologous and non-homologous vectors. The method described not only opens up opportunities for the genetic manipulation of C. gloeosporioides f. sp. aeschynomene, but also provides a framework for the development and optimization of transformation in other fungi.

Key words Electroporation Fungal transformation GFP Mycoherbicides 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • Micah Robinson
    • 1
  • Amir Sharon
    • 1
  1. 1.Institute of Cereal Crop Improvement, Department of Plant Sciences, Tel Aviv University, Tel Aviv 69978, Israel e-mail: a55555@post.tau.ac.il Tel.: +972-3-6406741 Fax: +972-3-6409380IL

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